Quantitative Determination of Zebularine (NSC 309132), a DNA Methyltransferase Inhibitor, and Three Metabolites in Murine Plasma by High-Performance Liquid Chromatography Coupled With On-Line Radioactivity Detection

J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Feb 2;831(1-2):147-55. doi: 10.1016/j.jchromb.2005.11.048. Epub 2005 Dec 20.

Abstract

The metabolism of zebularine (NSC 309132), a novel agent that inhibits DNA methyltransferases, is still uncharacterized. To examine the in vivo metabolism of zebularine, an analytical method was developed and validated (based on FDA guidelines) to quantitate 2-[(14)C]-zebularine and its major metabolites in murine plasma. Zebularine and its metabolites uridine, uracil and dihydrouracil were baseline-separated based on hydrophilic interaction chromatography by using an amino column. The assay was accurate and precise in the concentration ranges of 5.0-100 microg/mL for zebularine, 2.5-50 microg/mL for uridine, 1.0-10 microg/mL for uracil and 0.5-5.0 microg/mL for dihydrouracil. This assay is being used to quantitate zebularine and its metabolites in ongoing pharmacokinetic studies of zebularine.

Publication types

  • Research Support, N.I.H., Extramural
  • Validation Study

MeSH terms

  • Animals
  • Carbon Radioisotopes
  • Chromatography, High Pressure Liquid / methods*
  • Cytidine / analogs & derivatives*
  • Cytidine / blood
  • Cytidine / metabolism
  • DNA Modification Methylases / antagonists & inhibitors*
  • Drug Stability
  • Male
  • Mice
  • Mice, Inbred Strains
  • Online Systems
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Specific Pathogen-Free Organisms
  • Spectrophotometry, Ultraviolet
  • Uracil / analogs & derivatives
  • Uracil / blood
  • Uridine / blood

Substances

  • Carbon Radioisotopes
  • dihydrouracil
  • Uracil
  • Cytidine
  • pyrimidin-2-one beta-ribofuranoside
  • DNA Modification Methylases
  • Uridine